LOCUS       sequence-345687-        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAAV plasmid expressing an mCherry.3xFLAG.NOS1AP396-427 fusion 
            protein under the hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  AUTHORS   Freudenberg F, Candemir E, Chen X, Li LL, Esen-Sehir D, Schenk N, 
            Kinoshita M, Grunewald L, Frerichs V, Fattakhov N, Manchen J, Bikas 
            S, Kumar A, OLeary A, Slattery DA, von Engelhardt J, Courtney MJ, 
            Reif A
  TITLE     Hippocampal overexpression of NOS1AP promotes endophenotypes related
            to mental disorders.
  JOURNAL   EBioMedicine. 2021 Aug 26;71:103565. doi: 
            10.1016/j.ebiom.2021.103565.
  PUBMED    34455393
REFERENCE   2  (bases 1 to 5431)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "EBioMedicine."; date: "2021-08-26"; volume: "71:103565. doi"; 
            pages: " 10.1016/j.ebiom.2021.103565"
COMMENT     Sequence Label: pAAV-hSyn-mCherry.3xFLAG.NOS1AP396-427-WPRE
FEATURES             Location/Qualifiers
     source          1..5431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      480..489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             486..1193
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(631..649)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     899..918
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1200..1265
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1384..1972
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1437..1457)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1855..1866)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2004..2480
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2203..2222)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2520..2660
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2520..2649
                     /label=AAV2 ITR
     rep_origin      2735..3190
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2822..2841)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3032..3053
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3207..3226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3326..3348
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3386..3404)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3472..3576
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3577..3645,3646..4437)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3795..3814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4608..5196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5097..5116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5258..5387
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac cggatcctct agagtcgacg
      481 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
      541 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      601 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
      661 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
      721 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
      781 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      841 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
      901 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
      961 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     1021 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1081 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1141 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aaggaattcg
     1201 actacaaaga ccatgacggt gattataaag atcatgatat tgattacaag gatgacgatg
     1261 acaagctagc tcactcacca ctgctgggcg ctggcttggc tgactttgcc cacccagcgg
     1321 gcagcccttt aggtaggcat gactgcttgg tgaagctgga gtgcttttag taagcttatc
     1381 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1441 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1501 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     1561 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     1621 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     1681 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     1741 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     1801 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     1861 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     1921 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     1981 cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     2041 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     2101 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     2161 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     2221 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     2281 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     2341 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     2401 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     2461 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     2521 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2581 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2641 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2701 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     2761 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     2821 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2881 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2941 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3001 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3061 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     3121 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3181 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3241 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3301 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3361 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     3421 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3481 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3541 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3601 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3661 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3721 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3781 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3841 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3901 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3961 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4021 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4081 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4141 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4201 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4261 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4321 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4381 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4441 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4501 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4561 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4621 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4681 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4741 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4801 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4861 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4921 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4981 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5041 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5101 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5161 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5221 cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc
     5281 tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     5341 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg gccgcacgcg
     5401 tgtgtctaga ctgcagaggg ccctgcgtat g
//
